HK1241736B - Formulations of a pi3k/mtor-inhibitor for intravenous administration - Google Patents

Description

Formulations of PI3K/MTOR-inhibitors for intravenous administration

The present invention relates to a pharmaceutical formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or a pharmaceutically acceptable salt thereof. More particularly, the present invention relates to an aqueous pharmaceutical formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or a pharmaceutically acceptable salt thereof, wherein the aqueous pharmaceutical formulation is a clear solution. Such a formulation is particularly suitable for intravenous administration to a patient.

1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea and its preparation are disclosed in WO2009/143313. This compound is an inhibitor of PI3 kinase and mTOR and can be used to treat cancer.

Crystalline forms of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea and methods for their preparation are disclosed in WO 2010/096619.

1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea has the following chemical structure:

1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea can be prepared in a crystalline form and is chemically and physically stable in this form for up to 3 years at 25°C and 60% relative humidity (RH). However, the free base is not sufficiently water soluble to prepare aqueous solution formulations at therapeutic dose levels suitable for intravenous or parenteral administration.

There is a need to develop pharmaceutically acceptable formulations of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea that (a) are chemically stable upon storage (e.g., at 25° C. and 60% RH), and/or (b) will facilitate effective intravenous (or parenteral) administration of the drug to mammals, including humans.

Preferably, the formulation is suitable for intravenous administration of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, given its particular pharmacokinetic and bioavailability characteristics.

It is important that any intravenous formulation of a drug is a solution to facilitate safe and effective administration to the patient. It must be free of particles and not form a gel or suspension. Clear aqueous solutions are preferred.

A clear solution is defined as a visually clear solution that is substantially free of any visible particulate matter that can be observed upon visual inspection. Generally, if any particulate matter is observed, the formulation is not suitable for intravenous administration and should not be used due to the potential for vascular occlusion. Therefore, given the qualitative nature of the visual test, the term "substantially free of any visible particulate matter" is generally applied when no visible particulate matter is observed.

Particulate matter can be defined as follows:

Specks - discrete particles whose shape cannot be determined without magnification

Smoke or vortex - fine particles that look like smoke or a tornado and usually originate from the bottom of the sample bottle and swirl upward as the bottle rotates

Flocculated material - loosely aggregated particles or soft flakes

• Particles with a definite shape or characteristics that can be described as glassy, metallic, etc.

This visual inspection can be performed according to the method defined in European Pharmacopoeia method 2.9.20 entitled "Particulate contamination: visible particles". This method determines particulate contamination of injections and infusions by the presence of foreign, mobile, undissolved particles other than air bubbles in the solution. This test is intended to provide a simple procedure for visually assessing the quality of parenteral solutions with respect to visible particles. Other validated methods may also be used.

In the European Pharmacopoeia method 2.9.20, the apparatus (see "Figure 2.9.20.-1" shown in Figure 2) comprises an observation station containing:

A matte black board of appropriate size held in a vertical position

A non-glare white board of appropriate size held in a vertical position next to the black board

An adjustable lamp holder fitted with a suitable shaded white light source and a suitable light diffuser (a viewing illuminator containing two 13-watt fluorescent tubes, each 525 mm long, is suitable). The illumination intensity at the observation point is maintained between 2000 lux and 3750 lux, with higher values preferred for colored glass and plastic containers.

The method states: "Remove any adhered labels from the container and wash and dry the outside. Gently turn or invert the container, making sure not to introduce air bubbles, and observe in front of a white plate for approximately 5 seconds. Repeat the process in front of a black plate. Record the presence of any particles."

It has now been unexpectedly discovered that the technical problem is solved by the following aqueous pharmaceutical preparation, which comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or a lactate thereof, lactic acid and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 6 mg/ml and there is sufficient lactic acid to provide a clear solution; or 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or a phosphate salt thereof, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml and sufficient orthophosphoric acid is present to provide a clear solution (hereinafter referred to as the "formulation of the present invention").

Such a formulation can be administered directly to a patient intravenously or parenterally (to avoid degradation), preferably with the addition of a tonicity modifier. Alternatively, for administration to a patient at a later date, such a formulation, optionally containing a bulking agent and/or a tonicity modifier, can be freeze-dried to produce a freeze-dried solid composition that is chemically stable upon storage, preferably for at least 2 years, and then the freeze-dried solid composition can be constituted or reconstituted to provide a clear aqueous solution, preferably with the addition of a tonicity modifier, as needed, before administration to a patient via an intravenous (or parenteral) route.

It has been found that the alternative acids to lactic acid or orthophosphoric acid used in the formulations of the present invention, at preferred concentrations of 2.5 to 5.5 mg/ml of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, result in cloudy formulations containing particulate matter or gelling, and do not form the substantially clear, particle-free solutions desired for intravenous (or parenteral) administration to patients.

Regarding preparations containing lactic acid:

It has been found that at concentrations of 6 mg/ml or greater of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea solution, the necessary clear solution is not obtained or is not consistently obtained at the pH required for intravenous administration to patients.

Preferably, the present invention provides an aqueous pharmaceutical solution formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 6 mg/ml and sufficient lactic acid is present to provide a clear solution.

The concentration of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea in the formulations of the present invention may be 1 to 5.5 mg/ml, 2 to 5.5 mg/ml, or 3 to 5.5 mg/ml (calculated as the named free base).

Preferably, the present invention provides an aqueous pharmaceutical solution formulation, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of 2.5 to 5.5 mg/ml.

Preferably, the present invention provides an aqueous pharmaceutical formulation wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml.

Preferably, when the free base of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is used, greater than 2.5 molar equivalents of lactic acid are present in the formulation of the present invention. More preferably, 3 to 10, greater than 2.5 to 8.0, or 3.5 to 4.5 molar equivalents of lactic acid are present in the formulation of the present invention. Most preferably, about 4.1 molar equivalents of lactic acid are present in the formulation of the present invention.

Preferably, when the free base of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is used, the present invention provides an aqueous pharmaceutical solution formulation wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of 5.0 to 5.5 mg/ml and at least 2.5 molar equivalents of lactic acid are present.

Preferably, when the free base of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is used, the present invention provides an aqueous pharmaceutical solution formulation wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml and at least 2.5 molar equivalents of lactic acid are present.

Preferably, when the free base of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is used, the present invention provides an aqueous pharmaceutical solution formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml and lactic acid is present in an amount of at least 2.5 molar equivalents and sufficient to ensure a clear solution.

It should be noted that 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea forms a 1:1 (molar equivalent) lactate salt with lactic acid. The formulations of the present invention can be prepared using either the free base of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea or the lactate salt of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea. When lactate is used, it is preferred to use more than 1.5 molar equivalents of lactic acid to obtain a preferred lower limit of more than 2.5 molar equivalents of lactic acid present in the formulation of the invention.

Preferably, the present invention provides an aqueous pharmaceutical solution formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea lactate, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml and lactic acid is present in an amount of at least 1.5 molar equivalents and sufficient to ensure a clear solution.

DL-lactic acid, D-lactic acid or L-lactic acid, or any combination thereof can be used in the formulation of the present invention. Preferably, DL-lactic acid is used.

Preferably, the pH of the formulation of the present invention is not greater than 3.7. More preferably, the pH of the formulation of the present invention is 3.0 to 3.7, 3.3 to 3.6, or 3.4 to 3.5.

In a preferred embodiment, the present invention provides an aqueous pharmaceutical solution formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of up to 5.5 mg/ml and lactic acid is present in an amount greater than 2.5 molar equivalents and sufficient to ensure the formation of a clear solution having a pH no greater than 3.7.

In a preferred embodiment, the present invention provides an aqueous pharmaceutical solution formulation comprising 1-(4-([4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml and lactic acid is present in an amount of about 4.1 molar equivalents sufficient to ensure a clear solution having a pH no greater than 3.7.

A crystalline form of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea L-lactate can be used to prepare the formulations of the present invention. Preferably, the crystalline form of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea L-lactate used has a PXRD pattern (measured using a Bruker D4 diffractometer and copper K-alpha radiation) having major peaks at approximately 16.2, 17.3, 18.4, 18.9, 19.9, 20.9, and 23.1° 2θ (+/- 0.2° 2θ). The crystalline form of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea L-lactate is distinguished from other known forms of the salt by characteristic peaks at approximately 6.5, 15.9, 20.9, 22.1, and 23.1° 2θ (+/- 0.2° 2θ).

Regarding preparations containing orthophosphoric acid:

Preferably, the present invention provides an aqueous pharmaceutical solution formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml and sufficient orthophosphoric acid is present to provide a clear solution.

Preferably, the present invention provides an aqueous pharmaceutical solution formulation, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of 3.0 to 3.5 mg/ml.

• Preferably, the present invention provides an aqueous pharmaceutical formulation wherein at least 5 molar equivalents of orthophosphoric acid are used.

• Preferably, the present invention provides an aqueous pharmaceutical solution formulation wherein 5 to 7 molar equivalents of orthophosphoric acid are used.

In a preferred embodiment, the present invention provides an aqueous pharmaceutical formulation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml, and orthophosphoric acid is present in an amount of 5 to 7 molar equivalents in an amount sufficient to ensure a clear solution.

Preferably, the pH of the prepared formulation before intravenous administration is 2 to 2.5. The pH is then preferably adjusted to 3.0 to 4.5 for intravenous administration.

If the phosphate salt of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is used, it is preferably prepared using orthophosphoric acid.

If the formulation of the present invention is to be freeze-dried to provide a lyophilized solid composition, a filler is preferably added to the formulation before the freeze-drying process begins. The main function of the filler is to provide a structurally intact freeze-dried solid that does not collapse, which allows rapid reconstitution when forming the aqueous formulation before administration, and which should also promote effective lyophilization. Fillers are usually used when the total mass of solutes in the formulation is less than 2g/100ml. Fillers can also be added to obtain isotonicity with blood. The filler can be selected from a carbohydrate, a sugar alcohol, an amino acid or a polymer, or a mixture of any two or more thereof. Preferably, the filler is a sugar or a sugar alcohol, or a mixture thereof. Preferably, the sugar is sucrose. Preferably, the sugar alcohol is mannitol.

Reconstitution of the lyophilized solid composition can be achieved by adding the necessary amount of water present before lyophilization to obtain a clear solution. A tonicity adjuster can then be added before use.

Formulation of the lyophilized solid composition can be achieved by using an appropriate amount of water and/or an aqueous solution of a suitable tonicity adjusting agent to ensure a clear solution.

A tonicity adjusting agent must be present before the formulation is administered intravenously or parenterally to a patient by injection to avoid shrinkage or hemolysis of red blood cells and to alleviate or avoid pain and discomfort in the patient. This requires that the formulation to be administered to a patient have an effective osmotic pressure approximately the same as that of the patient's blood.

Suitable tonicity regulators are nonionic tonicity regulators, such as glycerol, sorbitol, mannitol, sucrose, propylene glycol or glucose, or a mixture of any two or more thereof. Preferably, the nonionic tonicity regulator is glucose, sucrose or mannitol, or a mixture of any two or more thereof.

Aqueous pharmaceutical formulations suitable for intravenous administration typically have a pH of 3 to 9. Formulations of the invention to be administered intravenously preferably have a pH of 3 to 4.5.

The formulations of the present invention can be used for radical treatment, palliative treatment, or preventive treatment of cancer in mammals, including humans. The cancer to be treated can be selected from the group consisting of leukemia, skin cancer, bladder cancer, breast cancer, uterine cancer, ovarian cancer, prostate cancer, lung cancer, colon cancer, pancreatic cancer, kidney cancer, stomach cancer, and brain cancer.

The weekly dosage of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea to be administered by the intravenous route for the treatment of cancer using the formulations disclosed herein is preferably in the range of 100 to 400 mg/ml per week.

The following examples illustrate the preparation of formulations of the present invention.

[Example]

Example 1

Prepare a 5 mg/ml solution of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimethylamino)piperidin-1-yl]carbonyl}phenyl]- Aqueous pharmaceutical preparation of morpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea and DL-lactic acid

D, L-lactic acid (334 mg) was dissolved in water for infusion to prepare a solution with a total volume of 100 ml. 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazine-2-yl)phenyl]urea (200 mg) was dissolved in 37 ml of the lactic acid solution, mixed for 120 minutes using an Ultra Turrax T25 (trademark) homogenizer, and the solution was sonicated in an ultrasonic bath for 10 minutes. The mixture was then stirred overnight using a magnetic stirrer to provide a clear solution. A volumetric flask was used to make up the volume to 40 ml with lactic acid solution. The solution was filtered into a clean 50 ml vial using a 0.2 μm nylon filter in a laminar flow (LAF) cabinet. The first 5 ml of the filtered solution was used to wet the filter and discarded as a non-representative filtered solution. The vial was crimped and sealed using a clean lyo-stopper and flip-top cap. The solution was visually inspected and found to be a clear, colorless solution.

Example 2

Preparation of (a) a mixture containing 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholine- 5 mg/ml aqueous solution of 4-amino-1,3,5-triazin-2-yl)phenyl]urea, DL-lactic acid and mannitol; and (b) preparation of Freeze-dried solid composition thereof

(a) 36,100 g of water for injection were weighed into a container. 125.82 g of DL-lactic acid (90.6% purity, parenteral grade) was slowly added and the mixture was stirred until the lactic acid dissolved. 195.3 g of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea was slowly added and the mixture was stirred until the material dissolved. 1900 g of mannitol powder (parenteral) was gradually added and the mixture was stirred until the material dissolved. Water for injection was added to make the solution up to a total weight of 38,760 g and the solution was stirred for an additional 10 minutes. The pH was checked and found to be 3.4 and the solution temperature was 29.3° C. The solution was sterile filtered through an in-line 0.45 μm clarification filter and a 0.22 μm filter assembly. This solution was then filled into 50 mL vials with a target fill volume of 20.8 mL per vial.Each vial was partially stoppered (not sealed) with a 20 mm Gray Lyo D777-1 V10-F597W FluroTec Siliconised (Trade Mark) stopper.

(b) The vials were then loaded into stainless steel trays and inserted into an LSL1000 (trademark) freeze dryer. The shelf temperature was set to 5° C. A freeze drying cycle was performed using the following method.

Backfill the freeze dryer with sterile filtered nitrogen to a set point of approximately 700 mbar (70,000 Pa) and completely close the vials using stoppers. The freeze dryer is then vented to atmospheric pressure using sterile filtered air and the vials are removed from the freeze dryer.

Each vial contains the freeze-dried (lyophilized) formulation as a white solid.

Example 3

Reconstitution of a composition comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4- Preparation of a 5 mg/ml aqueous solution of (4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, DL-lactic acid and mannitol agent

A vial of the lyophilized solid composition sample prepared in Example 2(b) was reconstituted as follows.

Approximately 25 ml of water for injection was placed in a syringe and a 0.2 micron PVDF filter membrane was attached to the syringe. Approximately 5 ml of the water was filtered through the membrane and discarded. The remaining 20 ml of water in the syringe was then filtered into a 50 ml vial containing the lyophilized solid composition prepared as described in Example 2(b). The mixture was swirled in the vial until a clear, colorless solution was obtained.

The reconstituted solution was analyzed as follows:

(a) pH

The pH of the solution in the vial was measured at 23.2°C to be pH = 3.52.

(b) Visual appearance of the reconstituted solution

One of the reconstituted vials was visually inspected using a method based on the aforementioned European Pharmacopoeia method 2.9.20. This method was designed to observe the presence of any visible particles.

By this method, the solution in the vial was visually inspected using a 3250 lux photometer reading in a Verivide DCAC60 (trade mark) light box against a matte black and white panel.

The results showed that reconstitution resulted in a clear, colorless solution that was free of particulate matter.

(c) Analysis of subvisible particles

The HIAC apparatus (trademark) was used to assess the presence of subvisible particles in the vial solution using a subvisible particle count method based on the definition of the USP 36 <788> Method 1 ("Light Obscuration Particle Count Test"). Because the USP 36 <788> Method 1 standard for Test 1.B defines the widest possible acceptable limit for subvisible particulate matter, these results must comply with this standard in order for the solution to be suitable for parenteral or intravenous administration. This test is described as follows:

“Test 1.B (Solutions for Parenteral Infusion or Solutions for Injection Supplied in Containers with a Nominal Content of Less than 100 mL)—The preparation complies with this test if the average number of particles present in the unit tested is not more than 6000 particles of 10 μm or greater per container and not more than 600 particles of 25 μm or greater per container.”

Using this method, first, 10 vial solution samples were combined. Four samples of less than 5 mL each were removed from the combined solution, and for each sample, the number of particles equal to or greater than 10 μm and 25 μm was counted using a HIAC HRLD 400 (trademark) sensor. The results obtained for the first sample were omitted. For each of the remaining three samples, the average number of particles per container was calculated and compared to the requirements of USP 36 <788> Test 1.B. Each of these samples met the acceptance criteria of USP 36 <788> Test 1.B for solutions suitable for parenteral or intravenous administration.

Example 4

Preparation of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1, Crystalline form of 3,5-triazin-2-yl)phenyl]urea L-lactate

Preparation A:

1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea (52 mg) was weighed into a 2 ml vial. A 22 mg/ml solution of L-lactic acid in 98:2 v/v ethyl acetate:dimethylformamide (0.5 ml) was added to the vial. The slurry was stirred at approximately 23° C. for 24 hours. The slurry was then filtered through a 0.2 μm nylon centrifugal filter to isolate the crystalline title compound.

The product was analyzed by PXRD (see "Study 7" below) using a Bruker D4 (trademark) diffractometer and copper K-alpha radiation, and the pattern shown in FIG1 was obtained.

Preparation B:

1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea (52 mg) was weighed into a 2 ml vial. A solution of 22 mg/ml of L-lactic acid in 98:2 v/v ethyl acetate:dimethylformamide (0.5 ml) was added to the vial. The slurry was heated to 60°C at a rate of 5°C/min, held at 60°C for 20 minutes, then cooled to 5°C at 0.1°C/min and held at this temperature until separation (24 hours after the start of the heating step). The slurry was filtered through a 0.2 μm nylon centrifugal filter to isolate the crystalline title compound.

The product was analyzed by PXRD (see "Study 7" below) using a Bruker D4 diffractometer with copper K-alpha radiation and gave a pattern consistent with that shown in FIG1 .

The following studies were conducted in connection with the present invention.

Research

1. About 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6- Study on 3 mg/ml aqueous formulation of dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

process

Nine individual acidic buffer solutions were prepared as follows such that each acid was used in approximately 6.8 molar equivalents (unless otherwise specified therein):

(WFI = Water for Infusion) ( ^* 90% w/w DL-lactic acid in water)

Stability testing was performed using three approximately 3 mg/ml samples of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea for each of the acid buffers prepared above. These samples were prepared as follows by weighing the required amount into each vial, using a target weight of 15.45 mg of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea (due to 97.1% API activity):

Buffer number (see above) N＝1 N=2 N=3 1 15.38mg 15.52mg 15.35mg 2 15.65mg 15.40mg 15.28mg 3 15.69mg 15.36mg 15.47mg 4 15.67mg 15.48mg 15.41mg 5 15.54mg 15.33mg 15.58mg 6 15.93mg 15.35mg 15.50mg 7 15.74mg 15.33mg 15.33mg 8 15.79mg 15.75mg 15.33mg 9 15.36mg 15.58mg 15.42mg 10 15.33mg 15.43mg 15.32mg

5 mL of the respective buffer was introduced into the vials after weighing the sample, each vial was closed by crimping the cap and then sealed with a protective film.The vials were placed on a roller bed in an oven at 25°C for 5 days.

result

At the end of the 5 day period, the pH of each sample was measured and each sample was visually observed using a light box as described in European Pharmacopoeia Method 2.9.20 (above), examining the samples against black and white backgrounds. The samples were also tested by illuminating the test using a narrow (Tyndall) beam light source and then visually inspected from a direction perpendicular to the beam to identify undissolved solid particles.

Six vials were found to have fallen off the roller during the course of the experiment, meaning the exact time these vials were actually rolled is unknown. These samples are marked with an asterisk ( ^* ) in the "pH Results" and "Visual Observations" tables below.

pH results after 5 days at 25°C

Buffer No. N＝1 N=2 N=3 1 (citric acid) 3.02 3.01 3.01 2(Succinic acid) 3.14 3.14 3.15 3(acetic acid) 3.86 3.87 3.88 4(orthophosphoric acid) 2.01 2.07 2.04 5(Glycine) 6.45 6.71 6.51 6(Tartaric acid) 7(DL-lactic acid) 2.91 2.91 8(Maleic acid) 1.91 1.90 9(Malic acid) 2.71 2.70 10(DL-lactic acid) 3.07 3.21 3.27

Visual observation of samples after 5 days at 25°C

Buffer No. N＝1 N=2 N=3 1 (citric acid) turbid turbid turbid 2(Succinic acid) turbid turbid turbid 3(acetic acid) Not in solution Not in solution Not in solution 4(orthophosphoric acid) turbid clarify turbid 5(Glycine) Not in solution Not in solution Not in solution 6(Tartaric acid) 7(DL-lactic acid) clarify clarify 8(Maleic acid) turbid turbid 9(Malic acid) turbid turbid 10(DL-lactic acid) clarify clarify clarify

in conclusion

These results show that a 3 mg/ml sample containing 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea and DL-lactic acid produced a clear solution after 5 days at 25°C. All other samples, except one (the orthophosphoric acid - N=2 sample), failed to produce a clear solution. Thus, acids other than DL-lactic acid and orthophosphoric acid are unsuitable for preparing aqueous pharmaceutical formulations with the desired API (active pharmaceutical ingredient) concentration for intravenous administration to patients.

2. About 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6- Study on 3 mg/ml and 4 mg/ml aqueous formulations of dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

process

(a) Four individual acidic buffer solutions were prepared for the 3 mg/ml formulation as follows, using approximately 6.8 molar equivalents of the individual acids:

(WFI = Water for Infusion)

Stability testing was performed using three approximately 3 mg/ml samples of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea for each of the acid buffers prepared above. These samples were prepared as follows by weighing the required amount into each vial, using a target weight of 15.45 mg of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea (due to 97.1% API activity):

buffer N＝1 N=2 N=3 hydrochloric acid 15.71 15.34 15.95 (D)-Lactic acid 15.52 15.40 15.50 (L)-Lactic acid 15.22 15.80 15.30 Orthophosphoric acid 15.43mg 15.50mg 15.79mg

(b) Prepare the individual acidic buffer solutions for the 4 mg/ml formulation as follows, using approximately 5.1 molar equivalents of the individual acid (unless otherwise specified):

(WFI = Water for Infusion) ( ^* 90% w/w DL-lactic acid in water)

Stability testing was performed using three approximately 4 mg/ml samples of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea for each of the acid buffers prepared above. These samples were prepared as follows by weighing the required amount into each vial, using a target weight of 20.60 mg of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea (due to 97.1% API activity):

Buffer No. N＝1 N=2 N=3 1 20.53 21.09 20.71 2 20.72 20.54 20.87 3 20.55 20.79 20.43 4 20.91 20.97 20.81 5 20.07 20.73 20.59 6 20.97 20.29 20.14 7 20.95 20.65 20.45 8 20.93 20.21 20.81 9 20.29 20.25 20.48 10 20.37 20.66 20.50 11 20.90 20.18 20.52 12 20.53 20.43 20.75 13 20.79 20.69 20.44 14 20.55 20.87 20.55

For both formulations (a) and (b) above, 5 mL of the respective buffer was introduced into the weighed sample in the vial, and each vial was closed by crimping the cap and sealed with a protective film. These vials were placed on a roller bed in an oven at 25° C. for 5 days.

result

At the end of the 5 day period, the pH of each sample was measured and each sample was visually observed using a light box as described in European Pharmacopoeia Method 2.9.20 (above), examining the samples against black and white backgrounds. The samples were also tested by illuminating the test using a narrow (Tyndall) beam light source and then visually inspected from a direction perpendicular to the beam to identify undissolved solid particles.

pH after 5 days at 25°C

3mg/mL

buffer Initial pH N＝1 N=2 N=3 33.3 mM hydrochloric acid 1.51 1.56 1.56 1.55 33.3mM (D)-lactic acid 2.68 3.14 3.07 3.09 33.3mM (L)-lactic acid 2.72 3.17 3.13 3.14 33.3 mM orthophosphoric acid 1.87 2.09 2.18 2.18

4 mg/mL

buffer Initial pH N＝1 N=2 N=3 33.3 mM citric acid 2.98 3.07 3.09 3.08 33.3 mM succinate 2.79 3.19 3.17 3.15 33.3 mM acetic acid 3.35 4.07 4.09 4.08 33.3 mM orthophosphoric acid 1.87 2.09 2.06 2.05 33.3 mM tartaric acid 2.28 2.41 2.42 2.42 33.3 mM hydrochloric acid 1.51 1.63 1.62 1.65 43mM (DL)-lactic acid 2.53 3.07 3.05 3.04 3.5 mM maleic acid 2.55 4.70 4.66 4.66 3.5 mM malic acid 3.08 5.14 5.11 5.11 33.3mM (D)-lactic acid 2.68 3.27 3.31 3.34 33.3mM (L)-lactic acid 2.72 3.36 3.37 3.38 33.3mM (DL)-lactic acid 2.60 3.23 3.23 3.28 33.3 mM maleic acid 1.72 2.03 2.04 2.05 33.3 mM malic acid 2.30 2.70 2.74 2.73

Visual observation of samples after 5 days at 25°C

3mg/mL

buffer N＝1 N=2 N=3 33.3 mM hydrochloric acid Presence of turbidity Presence of turbidity Presence of turbidity 33.3mM (D)-lactic acid clarify clarify clarify 33.3mM (L)-lactic acid clarify clarify clarify 33.3 mM orthophosphoric acid clarify clarify clarify

4 mg/mL

in conclusion

These results show that 3 mg/ml and 4 mg/ml samples containing 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea and D-lactic acid, L-lactic acid, or DL-lactic acid gave clear solutions after 5 days at 25°C.

These results show that the 3 mg/ml sample containing 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea and phosphoric acid also gave a clear solution after 5 days at 25°C.

All other samples failed to give clear solutions, and acids other than DL-lactic acid, D-lactic acid, L-lactic acid, and orthophosphoric acid would not be suitable for preparing aqueous pharmaceutical formulations with the desired API concentration for intravenous administration to patients.

3. 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6- Study on aqueous preparations of dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea at different pH and concentrations

process

(a) Buffer solutions for preparing 3 mg/ml, 5 mg/ml, 5.5 mg/ml, 6 mg/ml, and 6.5 mg/ml formulations of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea were prepared according to the following calculation (WFI = Water for Infusion) ( ^* 90% w/w DL-lactic acid in water).

3 mg/mL: 0.9 molar equivalents on a 10 mL scale equals 4.39 mg of DL-lactic acid ^* in 10 mL of WFI. (1)

3 mg/mL: 2.25 molar equivalents on a 10 mL scale equals 10.98 mg of DL-lactic acid ^* in 10 mL of WFI. (2)

3 mg/mL: 3.7 molar equivalents on a 10 mL scale equals 17.99 mg of DL-lactic acid ^* in 10 mL of WFI. (3)

5 mg/mL: 0.9 molar equivalents on a 10 mL scale equals 7.32 mg of DL-lactic acid ^* in 10 mL of WFI. (4)

5 mg/mL: 2.25 molar equivalents on a 10 mL scale equals 18.29 mg of DL-lactic acid ^* in 10 mL of WFI. (5)

5 mg/mL: 3.7 molar equivalents on a 10 mL scale equal to 30 mg of DL-lactic acid ^* in 10 mL of WFI. (6)

5 mg/mL: 7.2 molar equivalents on a 10 mL scale equals 58.54 mg of DL-lactic acid ^* in 10 mL of WFI. (7)

5 mg/mL: 10.8 molar equivalents on a 10 mL scale equals 87.80 mg of DL-lactic acid ^* in 10 mL of WFI. (8)

5.5 mg/mL: 2.25 molar equivalents on a 10 mL scale equals 20.12 mg of DL-lactic acid ^* in 10 mL of WFI. (9)

5.5 mg/mL: 3.7 molar equivalents on a 10 mL scale equals 33 mg of DL-lactic acid ^* in 10 mL of WFI. (10)

5.5 mg/mL: 10.8 molar equivalents on a 10 mL scale equals 96.58 mg of DL-lactic acid ^* in 10 mL of WFI. (11)

6 mg/mL: 3.7 molar equivalents on a 10 mL scale equals 36 mg of DL-lactic acid ^* in 10 mL of WFI. (12)

6 mg/mL: 3.7 molar equivalents on a 20 mL scale equals 72 mg of DL-lactic acid ^* in 20 mL of WFI. (14)

6.5 mg/mL: 3.7 molar equivalents on a 10 mL scale equals 39 mg of DL-lactic acid ^* in 10 mL of WFI. (13)

6.5 mg/mL: 3.7 molar equivalents at 20 mL scale equals 78 mg of DL-lactic acid ^* in 10 mL of WFI. (15)

Prepare the buffer solution in a 10 ml ( ^* 20 ml where indicated in the table below) volumetric flask using WFI using the following DL-lactic acid weights ( ^** 90% w/w DL-lactic acid in water):

Buffer No. Actual molar equivalent pH 1 3.91 0.8 3.14 2 12.12 2.5 2.86 3 18.16 3.7 2.69 4 6.97 0.9 2.99 5 19.07 2.3 2.68 6 30.80 3.8 2.55 7 57.10 7.0 2.41 8 87.82 10.8 2.30 9 20.36 2.3 2.67 10 33.37 3.7 2.56 11 100.33 11.2 2.27 12 35.93 3.6 2.55 13 41.84 3.9 2.50 14 3.6 2.50 15 3.7 2.45

(b) Weigh the following amount of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea into a vial (note that 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea has an activity of 97.1% and the target weights below will be corrected for this activity).

5 mL of the corresponding DL-lactic acid buffer was added to the API in the vials, and then each vial was closed with a crimp cap and sealed with a protective film.

Samples 1 to 11 were placed on a roller bed at room temperature and 50 rpm for approximately 21.5 hours.

Samples 12 and 13 were placed on a roller bed at room temperature and 50 rpm for approximately 23 hours.

Samples 14 and 15 were placed on a roller bed at room temperature and 50 rpm for approximately 25 hours.

Results after approximately 21.5/23/25 hours

At the end of the specified rolling period, the pH of each sample was measured and each sample was visually observed using a light box as described in European Pharmacopoeia Method 2.9.20 (above), and the samples were examined against black and white backgrounds. The samples were also tested by illuminating the test using a narrow (Tyndall) beam light source and then visually inspected from a direction perpendicular to the beam to identify undissolved solid particles.

pH

Visual assessment

Make the following observations:

Samples 7, 8, and 11 showed that they became liquid within 2 hours of being placed on the roller bed.

Sample 6 showed a solution within 4 hours of being placed on the roller bed

• Samples 3 and 10 showed a solution within 20 hours of placement on the roller bed.

in conclusion

After a period of approximately 24 hours, it was concluded that to obtain a clear solution, the solution concentration of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea must be less than 6 mg/ml and more than 2.5 molar equivalents of DL-lactic acid must be used in the formulation. However, the results of Samples 14 and 15 above showed that clear solutions were obtained at both solution concentrations of 6 mg/ml and 6.5 mg/ml of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea at 3.6 and 3.7 molar equivalents of DL-lactic acid, respectively. When compared to the results of Samples 12 and 13, the results of Samples 14 and 15 reflect the fact that there may be a metastable zone where both clear and non-clear solutions may form.

Results after a 72-hour period

After the aforementioned approximately 21.5 hour rolling period, samples 1 to 11 were stored at room temperature and not rolled for an additional period of time, providing a total experimental period of approximately 72 hours. It was observed that some samples that were not in solution after the initial approximately 21.5 hour rolling period became in solution by the end of the total 72 hour period.

Following the aforementioned approximately 25 hour tumbling period, samples 14-15 were tumbling stored at room temperature for an additional period of time to provide a total experimental period of approximately 73 hours.

The samples were visually evaluated using a light box as described in Ph. Eur. Method 2.9.20 (above), examining the samples against black and white backgrounds. The samples were also tested by illuminating with a narrow (Tyndall) beam light source and then visually inspected perpendicular to the beam to identify undissolved solid particles. The pH was also measured. The results are as follows:

(i) Visual assessment after 72 hours

(ii) Comparison of pH results and visual assessment after a period of approximately 24 hours and 72 hours

in conclusion

After a total experimental period of 72 hours, it was concluded that clear solutions were obtained with solution concentrations of 5 and 5.5 mg/ml of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, wherein at least 2.3 molar equivalents of DL-lactic acid were used in the formulation. A clear solution was also obtained with a solution concentration of 3 mg/ml of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, wherein more than 2.5 molar equivalents of DL-lactic acid were used in the formulation. The results of Samples 14 and 15 above show that clear solutions can be obtained at 6 mg/ml and 6.5 mg/ml solution concentrations of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea at 3.6 and 3.7 molar equivalents of DL-lactic acid, respectively. When compared with the results of Samples 12 and 13, the results of Samples 14 and 15 reflect the possibility of a metastable region in which both clear and non-clear solutions can form.

4. Regarding the use of 6.8 molar equivalents of orthophosphoric acid in 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}benzene Study on 3 mg/ml aqueous formulation of 3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

process

Prepare approximately 33.3 mM orthophosphoric acid in water as follows: Dispense 0.32569 g of orthophosphoric acid into approximately 80 mL of water for infusion. Make up to 100 mL in a volumetric flask using water for infusion and record the pH as 1.92.

A concentration of 3 mg/mL of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is desired and this must be considered in view of the 97.1% potency.

A 10 mL scale was determined based on this, resulting in a target weight of 30.9 mg for 1-(4-([4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea. Three API samples were prepared in 20 mL vials using the following weights:

N＝1 30.68mg N=2 31.21mg N=3 31.04mg

10 mL of the orthophosphate buffer prepared above was dispensed into each vial using an air displacement pipette. Each of these vials was closed with a crimp-tight cap and sealed with a protective film.

The samples were placed on a roller bed at room temperature for approximately 19 hours.

The samples were visually evaluated using a light box as described in Ph. Eur. Method 2.9.20 (above), examining the samples against black and white backgrounds. The samples were also tested by illuminating with a narrow (Tyndall) beam light source and then visually inspected perpendicular to the beam to identify undissolved solid particles. The pH was also measured. The results are as follows:

Visual assessment Final pH N＝1 clarify 2.15 N=2 clarify 2.15 N=3 clarify 2.17

dilution

Although clear, particle-free solutions were obtained by the above methods, the pH of each sample was too low to be preferred for intravenous administration, the preferred pH being 3 to 4.5.

Therefore, these three samples were diluted to 0.5 mg/mL, 0.1 mg/mL, and 0.05 mg/mL to determine if the pH had increased to an appropriate pH for intravenous administration. The diluted samples were placed on a roller bed overnight to reach equilibrium. The pH was also measured. The pH of these samples was as follows:

0.5 mg/mL

N＝1 2.64 N=2 2.63 N=3 2.64

0.1mg/mL

0.05mg/mL

N＝1 3.47 N=2 3.48 N=3 3.49

Each sample was visually evaluated using a light box according to Ph. Eur. Method 2.9.20 (described above) by examining the sample against a black and white background. The sample was also tested by illuminating it with a narrow (Tyndall) beam light source and then visually inspecting it perpendicular to the beam to identify undissolved solid particles. Each sample was observed to be a visually clear solution.

in conclusion

These results demonstrate that it is possible to formulate a clear, particle-free 3 mg/ml aqueous solution of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea with 6.8 molar equivalents of orthophosphoric acid. However, the pH of this formulation or its reconstituted formulation is not suitable for intravenous administration and must therefore be subsequently diluted to below 0.5 mg/mL to achieve a solution pH suitable for intravenous administration.

When these results are compared with those obtained for the lactic acid formulations described above, the pH and 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea concentrations that can be achieved are lower when orthophosphoric acid is used. Therefore, lactic acid is generally more suitable than orthophosphoric acid for preparing aqueous solution formulations for intravenous administration of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea according to the present invention.

5. About 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimethylamino)piperidin-1-yl]carbonyl}phenyl]- Study on 3mg/ml, 4mg/ml and 5mg/ml aqueous formulations of morpholin-4-yl-1,3,5-triazin-2-ylphenyl]urea

process

To prepare a 33.3 mM acetic acid solution, 0.2071 g of glacial acetic acid was dispensed into a 250 mL glass beaker and approximately 80 mL of WFI (water for infusion) was added.

0.0138 g of sodium acetate trihydrate was added and dissolved into solution. The solution was made up to 100 mL volume in a volumetric flask using WFI and the pH was recorded as 3.35.

Three API concentrations are desired (3, 4, and 5 mg/mL), which must be corrected to account for an API potency of 97.1%.The API weight is determined according to the following calculation.

30mg of activity is 30.9mg of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

40mg of activity is 41.2mg of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

50mg of activity is 51.49mg 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

Dispense the following weights into 20 mL glass vials:

3mg/mL 4 mg/mL 5mg/mL N＝1 30.86mg 41.13mg 51.49mg N=2 30.85mg 41.12mg 51.55mg N=3 30.76mg 41.21mg 51.66mg

10 mL of the acetic acid buffer prepared above was introduced into each weighed sample. Each of these vials was closed with a crimp-tight cap and sealed with a protective film.

The samples were placed on a roller bed at room temperature and visually evaluated using a light box as described in Ph. Eur. Method 2.9.20 (above), examining the samples against black and white backgrounds. The samples were also illuminated using a narrow (Tyndall) beam light source and then visually inspected perpendicular to the beam to identify undissolved solid particles. Visual analysis was performed at 24 hours, 48 hours, 72 hours, and 6 days.

result

No samples gave clear solutions after any of these 24 hour, 48 hour, 72 hour and 6 day periods.

The pH of the samples was assessed as follows:

(a) pH check after 48 hours

Initial buffer pH = 3.35

(i) Regarding 3 mg/mL samples

N＝1 3.88 N=2 3.89 N=3 3.90

(ii) Regarding 4 mg/mL samples

N＝1 3.94 N=2 3.95 N=3 3.95

(iii) Regarding 5 mg/mL samples

N＝1 3.99 N=2 4.00 N=3 4.00

(b) pH check after 6 days

(i) Regarding 3 mg/mL samples

N＝1 3.99 N=2 3.95 N=3 3.96

(ii) Regarding 4 mg/mL samples

N＝1 4.07 N=2 4.03 N=3 4.04

(iii) Regarding 5 mg/mL samples

N＝1 4.18 N=2 4.18 N=3 4.19

in conclusion

The results showed that 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea did not produce clear solutions at concentrations of 3, 4, and 5 mg/ml using 33.3 mM acetic acid. The 3 mg/ml aqueous formulation used contained approximately 6.8 molar equivalents of acetic acid. The 4 mg/ml aqueous formulation used contained approximately 5.1 molar equivalents of acetic acid. The 5 mg/ml aqueous formulation used contained approximately 4.1 molar equivalents of acetic acid.

6. Regarding the use of orthophosphoric acid for 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6- Study on 3 and 3.5 mg/ml aqueous formulations of dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea

Prepare a 33.3 mM aqueous solution of orthophosphoric acid as follows: Dispense 0.32767 g of orthophosphoric acid into approximately 75 mL of water for infusion. Make up to 100 mL in a volumetric flask using water for infusion and record the pH as 1.94.

3 and 3.5 mg/mL formulations of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea are desired and this must be considered in view of the 97.1% potency.

The 3 mg/ml aqueous formulation used contained approximately 6.8 molar equivalents of orthophosphoric acid. The 3.5 mg/ml aqueous formulation used contained approximately 5.9 molar equivalents of orthophosphoric acid.

The 5 mL scale was determined accordingly, so the target weight of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea for the formulation was 15.5 mg for the 3 mg/mL formulation and 18.0 mg for the 3.5 mg/mL formulation. Three samples were prepared for each formulation in 20 mL vials using the following weights:

3mg/mL 3.5 mg/mL N＝1 15.48mg 18.32mg N=2 16.15mg 18.02mg N=3 15.89mg 18.28mg

5 mL of the orthophosphate buffer prepared above was dispensed into each vial using an air displacement pipette. Each of these vials was closed with a crimp-tight cap and sealed with a protective film.

The samples were placed on a roller bed at room temperature for 15 hours.

The samples were visually evaluated using a light box as described in European Pharmacopoeia Method 2.9.20 (above), examining the samples against black and white backgrounds. The samples were also tested by illuminating with a narrow (Tyndall) beam light source and then visually inspected perpendicular to the beam to identify undissolved solid particles. All solutions were observed to be visually clear. The pH was also measured.

The results are as follows (note that the entry pH of 33.3 mM orthophosphoric acid is pH = 1.94)

3mg/mL

N＝1 2.02 N=2 2.03 N=3 2.05

3.5 mg/mL

N＝1 2.09 N=2 2.07 N=3 2.07

in conclusion

These results show that it is possible to formulate clear, particle-free 3.0 or 3.5 mg/ml aqueous formulations of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea with 5.9 molar equivalents of orthophosphoric acid.

However, the pH readings indicate that dilution may be required to provide a suitable pH to allow direct intravenous or parenteral administration of these formulations.

When these results are compared to those obtained with the lactic acid formulations described above, lower pH and 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea concentrations were achieved when orthophosphoric acid was used. Therefore, lactic acid is preferred for the preparation of clear, particle-free aqueous formulations of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea suitable for intravenous or parenteral administration.

7.1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3, Characteristics of the crystalline form of 5-(5-triazin-2-yl)phenyl]urea L-lactate

PXRD analysis

Powder X-ray diffraction (PXRD) analysis was performed on a Bruker D4 (trademark) diffractometer using copper radiation (wavelength: ). The tube voltage and amperage were set to 35 kV and 40 mA, respectively. The divergence slit used was v6, and the scattering slit was set at 0.499 mm. A variable receiving slit was used. The diffracted radiation was detected by a Vantec detector. A θ-2θ continuous scan from 2.0 to 55° 2θ was used at 5.4°/min (0.2 sec/0.018° step). A corundum standard was analyzed to check instrument alignment. Data was collected and analyzed using Bruker AXS software. Samples were prepared by placing them on silicon wafers. PXRD spectra were visualized and evaluated using DIFFRAC.EVA V3.1 software. PXRD data files (.raw) were not processed before peak searching. Typically, a threshold of 1.3 and a width value of 0.3 were used for preliminary peak assignment. The output of the automated assignment was visually inspected to ensure validity and manual adjustments were made as needed. Additionally, peaks were manually assigned within the spectra where appropriate.The peak at 28.1° 2Θ related to the mounting medium was manually removed from this list.

X-ray diffraction measurements are performed using Bragg-Brentano geometry on a Bruker instrument used for the measurements reported in this invention, with the sample typically placed on a flat silicon plate. The sample powder is pressed against a glass slide or equivalent to ensure a random surface and appropriate sample height. The sample holder is then placed in the instrument. The incident X-ray beam is directed at the sample, initially at a small angle relative to the plane of the holder, and then moved through an arc with a continuously increasing angle between the incident beam and the plane of the holder. Measurement differences associated with these X-ray powder analyses are caused by various factors, including: (a) errors in sample preparation (e.g., sample height), (b) instrument errors (e.g., flat sample errors), (c) calibration errors, (d) operator errors (including errors in determining peak positions), and (e) material properties (e.g., errors in preferred orientation and transparency). Calibration errors and sample height errors often arise when all peaks move in the same direction. Small differences in sample height when using a flat holder will result in large shifts in PXRD peak positions. Systematic studies have shown that using a Shimadzu XRD-6000 in a typical Bragg-Brentano configuration, a sample height difference of 1 mm results in peak shifts of up to 1° 2θ (Chen et al.; J Pharmaceutical and Biomedical Analysis, 2001; 26, 63). These shifts can be identified from the X-ray diffraction pattern and can be eliminated by compensating for the shift (applying a systematic correction factor to all peak position values) or recalibrating the instrument. As described above, it is possible to correct measurements from various different machines by applying a systematic correction factor to bring the peak positions into line. Typically, this correction factor will bring the peak positions from the Bruker measurements into line with the expected peak positions and can be in the range of 0 to 0.2° 2θ.

The PXRD pattern of the crystalline form of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea L-lactate of Example 4 Preparation A is provided in FIG1 and is characterized by the following list of peaks expressed in °2θ (+/- 0.2°2θ) and relative intensities (≥ 2.5%) as measured on a Bruker D4 diffractometer using copper K-α (CuKα) radiation:

( ^* Relative intensities may vary depending on crystal size and morphology)

The crystalline form of the 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea L-lactate salt is distinguished from other known (semi-crystalline) forms of the salt by having characteristic peaks at approximately 6.5, 15.9, 20.9, 22.1, and 23.1° 2θ (+/- 0.2° 2θ).

7. Chemical stability of the freeze-dried solid preparation of the present invention

Samples of the lyophilized solid formulation prepared according to the method of Example 2 in 50 mL clear vials were analyzed for chemical degradation at various time points after storage at 25°C/60% relative humidity (RH) and 40°C/75% RH. Several samples were evaluated for each condition to ensure representative results at the selected time points.

These 40°C/75%RH samples were tested after 6 months.

The 25°C/60% RH samples were tested after 6 months, 12 months, 24 months and 36 months.

The chemical purity of the samples was tested using high performance liquid chromatography (HPLC) using the following method to measure any degradation during the testing period.

HPLC method

Solutions, samples, and standards for the HPLC method were prepared as follows:

Reference Standard: 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea with known potency

Diluent: acetonitrile/water (1:1 v/v).

Mobile phase A: 10 mM aqueous ammonium bicarbonate buffer solution, pH adjusted to 9.8 with aqueous ammonium hydroxide solution.

Mobile phase B: acetonitrile

Sample solvent: Add 3 mL of 0.1 N aqueous hydrochloric acid to a 1000 mL volumetric flask and dilute to the desired volume with diluent (acetonitrile/water, 1:1 v/v). Mix thoroughly.

NOTE: Larger or smaller volumes of solution can be prepared by using appropriate ratios of components.

Standard and check standard preparations:

Accurately prepare two approximately 2 mg/mL (+/- 10%) solutions of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea reference standards in the sample solvent and accurately record their concentrations. These are the standard and check standard solutions. Produce the standard and check standard preparations by accurately diluting these solutions with this diluent to a concentration of approximately 2 μg/mL of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea.

Sensitivity solution:

• Use this diluent to accurately dilute the standard preparation to a concentration of approximately 0.06 μg/mL of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea.

Sample preparation:

Two vials of lyophilized solid formulation of 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea (prepared according to the method of Example 2) were reconstituted by adding 20 mL of water to each vial, shaking the vial to dissolve the solid, and waiting for the bubbles to disappear. The solution was transferred to a 1000 mL volumetric flask. Each vial was rinsed at least twice with diluent, and the rinse solution was transferred to the volumetric flask. Diluted to the set volume with diluent.

Chromatographic conditions:

Liquid chromatography system – for example, a Waters 2695 (trademark) or Agilent 1100 (trademark) machine

Column: Waters Xbridge C18 (trademark), 15 cm x 4.6 mm, 3.5 μm or equivalent

Column temperature: 40°C

Injection volume: 20 μL

Flow rate: 1.0 mL/min

Detection: 303nm UV

Running time: 60 minutes

Mobile phase A

Mobile phase B

Linear gradient table:

Time (minutes) % Mobile phase A % mobile phase B 0 90 10 37 50 50 47 10 90 52 10 90 53 90 10 60 90 10

Notes

The HPLC machine was prepared by pumping mobile phase B through the column until a stable baseline was obtained (this typically took approximately 30 minutes).

Before running the injection sequence, re-equilibrate the chromatography system with mobile phase A (usually 10 to 15 minutes).

Before running samples, ensure that the system is suitable for injection of blank diluents, sensitivity solutions, and standard preparations using the chromatographic conditions described above.

The following criteria must be met during initial HPLC setup or after any major changes to the system: It is recommended to inject at least one conditioning blank before testing system suitability.

^* Average of all system suitability (reproducibility) injections used.

^** Refer to the United States Pharmacopoeia (USP) calculation equations for efficiency and peak asymmetry.

Inject a check standard preparation according to the above chromatographic conditions.The response factor of the check standard preparation (calculated from the area of the standard, standard weight, dilution factor and purity factor) must be within ±5% of the standard preparation.

After demonstrating system suitability, inject the blank solution, standard preparation, and prepared test sample according to the chromatographic conditions described above, followed by the standard preparation. It is recommended that no more than six test sample injections be performed between standard preparation injections. For each injection (standard and sample), measure the retention time and area of the 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea peak in each chromatogram. For each sample injection, also measure the retention time and peak area of any peaks present in the sample injection that are not present in the blank injection. If gradient artifacts are present, do not integrate. Compare the blank injection chromatogram to the sample chromatogram to determine which peak in the sample is associated with the blank and gradient artifact peaks. Calculate the % of degradant and report individual degradant peaks at or above 0.05%. Unknown degradants should be reported individually with their relative retention times. Known degradants should be reported individually by name.

The results are summarized in the table below.

Keywords

NMT = not more than.

NR = Not reported.

RRT = relative retention time

All % are w/w

Degradation product 1

Degradation product 2

Degradants 3, 4, 5 and 6

Each of these is characterized only by their RRT.

Degradants 3, 5, and 6 were identified as process-related impurities that did not alter stability and were therefore not reported at the 36-month time point.

in conclusion

The results showed that samples of the lyophilized solid formulation prepared according to the method of Example 2 in 50 mL clear vials were chemically stable for at least 36 months at 25°C/60% RH and at least 6 months at 40°C/75% RH.

Claims (24)

1. A freeze-dried formulation, which can be obtained by freeze-drying an aqueous drug solution formulation, said aqueous drug solution formulation comprising: 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or its lactate, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present in the aqueous solution of the drug at a concentration of 5.0 to 5.5 mg/ml, and at least 2.5 molar equivalents of lactic acid are present to provide a clear solution; or 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or its phosphate, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present in a solution concentration of less than 4 mg/ml, and at least 5 molar equivalents of orthophosphoric acid are used in the aqueous solution of the drug to provide a clear solution. 2. The freeze-dried formulation of claim 1, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, or its lactate, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present in the aqueous pharmaceutical preparation at a concentration of 5.0 to 5.5 mg/ml, and at least 2.5 molar equivalents of lactic acid are present to provide a clear solution. 3. The freeze-dried formulation of claim 2, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present in the aqueous pharmaceutical preparation at a concentration of 5.0 to 5.5 mg/ml, and at least 2.5 molar equivalents of lactic acid are present to provide a clear solution. 4. The freeze-dried formulation of claim 2, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present in the aqueous drug formulation at a solution concentration of about 5 mg/ml. 5. The freeze-dried formulation of claim 2, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml, and at least 2.5 molar equivalents of lactic acid are present, sufficient to ensure the formation of a clear solution. 6. The freeze-dried formulation of claim 1, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of about 5 mg/ml, and lactic acid is present in an amount of about 4.1 molar equivalents sufficient to ensure the formation of a clear solution with a pH not greater than 3.7. 7. The freeze-dried formulation of claim 1, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, lactic acid, and water, wherein the amount of lactic acid present is greater than 2.5 to 8.0 molar equivalents and sufficient to ensure the formation of a clear solution. 8. The freeze-dried formulation of claim 2, wherein DL-lactic acid, L-lactic acid or D-lactic acid is used in the aqueous pharmaceutical preparation. 9. The freeze-dried formulation of claim 8, wherein DL-lactic acid is used in the aqueous pharmaceutical preparation. 10. The freeze-dried formulation of claim 1, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea or a phosphate thereof, orthophosphoric acid and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml, and at least 5 molar equivalents of orthophosphoric acid are used in the aqueous pharmaceutical preparation to provide a clear solution. 11. The freeze-dried formulation of claim 10, wherein the aqueous pharmaceutical preparation comprises 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml, and at least 5 molar equivalents of orthophosphoric acid are used in the aqueous pharmaceutical preparation to provide a clear solution. 12. The freeze-dried formulation of claim 11, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present in the aqueous pharmaceutical preparation at a concentration of 3.0 to 3.5 mg/ml. 13. The freeze-dried formulation of claim 10, wherein 5 to 7 molar equivalents of orthophosphoric acid are used in the aqueous pharmaceutical preparation. 14. The freeze-dried formulation of any one of claims 1 to 13, further comprising a filler. 15. The freeze-dried formulation of claim 14, wherein the filler is mannitol. 16. A method for preparing a clear aqueous solution formulation of a drug, said method being used to reconstruct or constitute the freeze-dried formulation of any one of claims 1 to 15 by using water or an aqueous solution containing a tension modifier. 17. The method of claim 16, wherein the tension modifier is glucose, sucrose, or mannitol, or a mixture of any two or more thereof. 18. The method of claim 16, wherein the clarified aqueous solution formulation of the drug is adjusted as needed to have a pH suitable for intravenous or non-enteral administration. 19. The method of claim 18, wherein the pH is 3 to 4.5. 20. An aqueous pharmaceutical preparation comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea or a phosphate thereof, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml, and at least 5 molar equivalents of orthophosphoric acid are used in the aqueous pharmaceutical preparation to provide a clear solution. 21. The aqueous pharmaceutical preparation of claim 20, comprising 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea, orthophosphoric acid, and water, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of less than 4 mg/ml, and at least 5 molar equivalents of orthophosphoric acid are used in the aqueous pharmaceutical preparation to provide a clear solution. 22. The aqueous pharmaceutical preparation of claim 21, wherein 1-(4-{[4-(dimethylamino)piperidin-1-yl]carbonyl}phenyl)-3-[4-(4,6-dimorpholin-4-yl-1,3,5-triazin-2-yl)phenyl]urea is present at a solution concentration of 3.0 to 3.5 mg/ml. 23. The aqueous pharmaceutical preparation of claim 20, wherein 5 to 7 molar equivalents of orthophosphoric acid are used. 24. Use of the formulation of any one of claims 1 to 15 and 20 to 23 in the preparation of a medicament for treating cancer.